Accelerated Wright-Giemsa and May-Grünwald staining methods

Abstract

The present disclosure provides methods for carrying out Romanowsky-type stains, specifically Wright-Giemsa and May-Grünwald stains, quickly and efficiently. The methods greatly reduce the overall amount of time required to complete a Wright-Giemsa stain or a May-Grünwald stain of sufficient quality on a biological sample. The subject methods can be applied to both manual and automated staining procedures.

What is claimed is: 
     
       1. A computer-implemented method for performing a stain on one or more biological samples in an automated staining system, comprising:
 executing instructions stored on a nontransitory computer-readable medium that direct the automated staining system to: 
 apply the one or more biological samples to a substrate in a sample application subsystem; 
 contact the one or more biological samples with a fixation reagent bath comprising a fixation reagent by transferring the one or more biological samples from the sample application subsystem to the fixation reagent bath via a sample transfer subsystem, wherein the one or more biological samples is contacted with the fixation bath for a period of time of from 15 seconds to 45 seconds; and 
 contact the one or more biological samples with a staining reagent bath comprising a staining reagent by transferring the one or more biological samples from the fixation reagent bath to the staining reagent bath via the sample transfer subsystem, wherein the one or more biological samples is contacted with the staining reagent bath for a period of time of from 15 seconds to 60 seconds. 
 
     
     
       2. The method of  claim 1 , wherein the method further comprises contacting the one or more biological samples with one or more rinse reagent baths comprising a rinse reagent by transferring the one or more biological samples from the staining reagent bath to the one or more rinse reagent baths via the sample transfer subsystem. 
     
     
       3. The method of  claim 1 , wherein the staining reagent is a Write-Giemsa stain. 
     
     
       4. The method of  claim 1 , wherein the one or more biological samples is placed in the fixation reagent bath for a period of time ranging from about 25 seconds up to about 35 seconds. 
     
     
       5. The method of  claim 1 , wherein the fixation reagent is methanol. 
     
     
       6. The method of  claim 3 , wherein the Write-Giemsa stain comprises methanol. 
     
     
       7. The method of  claim 3 , wherein the one or more biological samples is placed in the Wright-Giemsa staining reagent bath for a period of time ranging from about 25 seconds up to about 45 seconds. 
     
     
       8. The method of  claim 2 , wherein the rinse reagent is a phosphate buffer that has a pH value ranging from about 5.0 up to about 9.0 pH units. 
     
     
       9. The method of  claim 8 , wherein the phosphate buffer has a pH value ranging from about 6.5 up to about 7.0 pH units. 
     
     
       10. The method of  claim 2 , wherein the one or more biological samples is placed in the one or more rinse reagent baths for a period of time ranging from about 105 seconds up to about 135 seconds. 
     
     
       11. A computer-implemented method for performing a stain on one or more biological samples in an automated staining system, comprising:
 executing instructions stored on a nontransitory computer-readable medium that direct the automated staining system to: 
 apply the one or more biological samples to a substrate in a sample application subsystem; 
 contact the one or more biological samples with a fixation reagent bath comprising a fixation reagent by transferring the one or more biological samples from the sample application subsystem to the fixation reagent bath via a sample transfer subsystem, wherein the one or more biological samples is contacted with the fixation reagent bath for a period of time of from 15 seconds to 45 seconds; and 
 contact the one or more biological samples with a staining reagent bath comprising a staining reagent by transferring the one or more biological samples from the fixation reagent bath to the staining reagent bath via the sample transfer subsystem, wherein the one or more biological samples is contacted with the staining reagent bath for a period of time of from 165 seconds to 195 seconds. 
 
     
     
       12. The method of  claim 11 , wherein the method further comprises contacting the one or more biological samples with one or more rinse reagent baths comprising a rinse reagent by transferring the one or more biological samples from the staining reagent bath to the one or more rinse reagent baths via the sample transfer subsystem. 
     
     
       13. The method of  claim 11 , wherein the staining reagent is a May-Grünwald stain. 
     
     
       14. The method of  claim 13 , wherein the one or more biological samples is placed in the May-Grünwald staining reagent bath for a period of time ranging from about 175 seconds up to about 185 seconds. 
     
     
       15. The method of  claim 13 , wherein the May-Grünwald stain comprises methanol. 
     
     
       16. The method of  claim 12 , wherein the one or more biological samples is placed in the one or more rinse reagent baths for a period of time ranging from about 105 seconds up to about 135 seconds. 
     
     
       17. The method of  claim 12 , wherein the one or more biological samples is placed in the one or more rinse reagent baths for a period of time ranging from about 145 seconds up to about 155 seconds. 
     
     
       18. The method of  claim 12 , wherein the rinse reagent is a phosphate buffer that has a pH value ranging from about 5.0 up to about 9.0 pH units. 
     
     
       19. The method of  claim 18 , wherein the phosphate buffer has a pH value ranging from about 6.5 up to about 7.0 pH units.